Structure of selenocysteine synthase from Escherichia coli and location of tRNA in the seryl-tRNAsec–enzyme complex

Selenocysteine synthase of Escherichia coli catalyses the biosynthesis of selenocysteine in the form of the aminoacyl-tRNA complex, the reaction intermediate being aminoacrylyl-tRNA(sec) covalently bound to the prosthetic group of the enzyme. Selenocysteine synthase and the specific aminoacrylyl-tRNA(sec)-enzyme complex as well as the isolated seryl-tRNA(sec) were investigated in the electron microscope and analysed by means of image processing to a resolution of 2 nm in projection. The stoichiometric composition of the selenocysteine synthase molecule was elucidated by scanning transmission electron microscopic mass determination. The enzyme has a fivefold symmetric structure and consists of 10 monomers arranged in two rings. The tRNA is bound near the margin of the dimeric subunits. Principal component analysis of the tRNA-enzyme complexes revealed that the selenocysteine synthase appears to bind only one seryl-tRNA(sec) per dimer, which is consistent with the result of biochemical binding studies.     

The ecological value of Eryngium horridum in maintaining biodiversity in subtropical grasslands

The role of facilitation in the structuring of plant communities has been often demonstrated in environments under high abiotic stress, especially in semi‐arid and arid ecosystems and high elevations. Few studies, however, analysed facilitation in systems that are highly productive and rich in species, which are thought to be theoretically unlikely to demonstrate strong effects of facilitation. Here, we investigate the importance of Eryngium horridum, a rosette species, on the maintenance of plant diversity in subtropical grasslands in southern Brazil. We evaluated facilitation in areas under two different types of management: abandonment and grazing. Plots were established in areas with and without individuals of E. horridum and all species were identified and had their cover estimated. The Relative Neighbour Effect index was calculated in order to verify the presence of competition or facilitation. Our results indicated facilitation in both abandoned and in grazed grasslands, but apparently through different mechanisms. In the first case, the plant's architecture opens the canopy and allows more light to reach small forbs in the grass matrix. In the second case, E. horridum appears to protect more palatable species from herbivores. Otherwise considered an obnoxious species, E. horridum plays an important ecological role in subtropical grasslands in southern Brazil by facilitating other species and consequently, increasing local richness. Areas with this rosette species are important sources of diaspores, which are able to colonize new open sites and thus, maintain biodiversity.     

An Arabidopsis gene homologous to mammalian and insect genes encoding the largest proteasome subunit

A gene encoding a protein with extensive homology to the largest subunit of the multicatalytic proteinase complex (proteasome) has been identified in Arabidopsis thaliana. This gene, referred to as AtPSM30, is entirely encompassed within a previously characterized radiation-induced deletion, which may thus provide the first example of a proteasome null mutation in a higher eukaryote. However, the growth rate and fertility of Arabidopsis plants do not appear to be significantly affected by this mutation, even though disruption experiments in yeast have shown that most proteasome subunits are essential. Analysis of mRNA levels in developing seedlings and mature plants indicates that expression of AtPSM30 is differentially regulated during development and is slightly induced in response to stress, as has been observed for proteasome genes in yeast, Drosophila, and mammals. Southern blot analysis indicates that the Arabidopsis genome contains numerous sequences closely related to AtPSM30, consistent with recent reports of at least two other proteasome genes in Arabidopsis. A comparison of the deduced amino acid sequences for all proteasome genes reported to date suggests that multiple proteasome subunits evolved in eukaryotes prior to the divergence of plants and animals.GenBank accession number: M98495     

Analysis of Chlorophyll Fluorescence by Means of Noisy Light

Linearization is an efficient means of detecting individualcomponents in complex fluorescence induction curves. In a previousinvestigation based on sine-waves (Hansen, Kolbowski, and Dau,1987) the components related to the plastoquinone pool, thehigh-energy state of the thylakoid membrane and the state 1-state2 transition controller could be identified. In this paper,binary noise is used as an alternative input signal which canalso allow linearization by mathematical tools. Comparison withexperiments using sine-waves shows that curve-fitting of thenoise experiments yields the same data as the sine-wave analysis.Further, some additional components were revealed as labelledby the related time-constants whose values depended on lightintensity (e.g. 5, 15, 20, 70, 85, 550 s for spinach at 2.5W m^–2). The advantages of noise analysis are a shorter measuring time and the availability ofan on-line criterion which indicates when a sufficient signal-to-noiseratio is reached. Key words: Chlorophyll fluorescence, correlation functions, linearization, noise, spinach, time-constants     

Relative importance of bacteria, microalgae and yeast for growth of the sponge Halichondria melanadocia (De Laubenfels, 1936): A laboratory study

Bacteria, microalgae and yeast less than 10 μm in size are the primary food source of sponges but their relative contribution to somatic growth is poorly understood. In a laboratory study, the sponge Halichondria melanadocia was fed for 6 weeks a diet consisting solely of four bacterial strains, or a mixed diet consisting of bacteria, microalgae and yeast. Both diets were fed at three concentrations, based on the natural concentration (NC) of particles available to sponges: 1/5, 1 and 5NC. Mean final size of H. melanadocia was 40% greater on a mixed diet than on the bacteria diet, probably because of the greater supply of carbon and other essential nutrients in microalgae and yeast. Cell concentration also significantly affected the growth of H. melanadocia, with greatest growth for sponges fed at the highest cell concentration. The estimated carbon requirement for H. melanadocia to meet metabolic costs was 0.356 mg C l^− 1 or 103 μg C h^− 1 gDW^− 1. Many H. melanadocia appeared to be optimizing their surface area for food uptake.     

DOPA synthesis in non-producer cultures of Mucuna deeringiana

Cell cultures of Mucuna deeringiana on various media fail to accumulate DOPA or stizolobic acid. Radiotracer studies prove that both compounds are being synthesized in the cells; however, the apparent lack of production is due to metabolic emphasis on catabolism instead of storage.     

Changes in phenotypic expression in embryonic and adult cells treated with 5-azacytidine

We have previously shown that 5-azacytidine (5-Aza-CR) induced the formation of biochemically differentiated myotubes, adipocytes, and chondrocytes in the mouse embryo cell line, C3H/10T1/2CL8 (10T1/2), and that the induction of the muscle phenotype was cell cycle specific. Here we show that the adipocyte phenotype is also induced maximally in cells treated during early S phase. During this period, the minimum treatment time required for the subsequent formation of myotubes was 5 min and the number of myotubes formed was dependent on treatment time. The incorporation of ¹⁴C-5-Aza-CR into DNA during the cell cycle, however, was not enhanced during early S phase, suggesting that incorporation of 5-Aza-CR into specific DNA sequences synthesized during early S phase may be required for the expression of the new phenotypes. Single cells, obtained by plating cell suspensions into 16 mm wells at limiting dilution, were treated with 5-Aza-CR during S phase. The resulting clones showed a high frequency of phenotypic conversion, indicating that 5-Aza-CR did not act via a selective mechanism, and several of the clones were capable of expressing more than one phenotype. The cells required more than 2 division cycles after treatment with the analog for the expression of the muscle phenotype and the capacity to differentiate was retained for long periods of time in the absence of cell division. The adult mouse line, CVP3SC6, differentiated into functional striated muscle cells following treatment with 5-Aza-CR. The analog also caused oncogenic transformation in the adult line at the same concentration that was effective at inducing myogenic expression.